• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.189-203
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• 1999

To determine sex of bovine embryos using hamster histocompatibility Y(H-Y) antibodies, bovine compact morulae were incubated for 6 hours in TCM199 supplemented with 10% hamster H-Y antiserum and the embryos with developmental arrest were diagnosed as male embryos, while the embryos showing development during the incubation as female embryos. This presumptive embryo sexing was confirmed by polymerase chain reaction(PCR)method. 1. In the result of hamster sperm cytotoxicity test to measure H-Y antibody titer, the rate of dead sperm was considerably lower in H-Y antiserum absorbed with hamster male splenocytes than in H-Y antiserum absorbed with hamster female splenocytes or H-Y antiserum unabsorbed with splenocytes(p<0.01). 2. The rate of oocytes fertilized in vitro and the rate of blastocysts of the fertilized oocytes were 58.5% and 32.4%, respectively. The rate of blastocysts on day 8 was 15.9%, denoting the highest rate during whole culture period posterior to in vitro fertilization (IVF). 3. The bovine 16 cell and compact morulae embryos incubated in the medium supplemented with hamster H-Y antibodies showed 37.1% and 48.9% of developmental arrest which were diagnosed as male, respectively, and rates of redeveloped embryos from the arrested were 24.1% in 16 cell and 44.3% in compact morulae embryos, respectively, denoting higher rate of sex determination and rate of redevelopment in compact morulae than 16 cell embryos. 4. Bovine compact morulae of Korean cattle and Holstein were treated with hamster H-Y antibodies for sex determination and the rates of developmental arrest(diagnosed as male) were 48.4% for Korean cattle and 47.9% for Holstein, respectively. The rates of redeveloped embryos to blastocyst after treatment were 42.6% for Korean cattle and 41.8% for Holstein, respectively, showing no significant differences of sex determination and redevelopment between both breed. 5. The sex determination of bovine embryos(Korean cattle and Holstein) using hamster H-Y antibodies was diagnosed by PCR for confirmation, denoting the rates of 86.1% for Korean cattle and 85.9% for Holstein male embryos, respectively, and the rates of 91.9% for Korean cattle and 90.1% for Holstein female embryos, respectively, with no significant differences of sex determination between both breed. These results indicated that hamster H-Y antibodies can be usable for sex determination of bovine embryos of Korean cattle and Holstein, the viability of bovine embryos was sustained while being cultured in the medium supplemented with hamster H-Y antibodies of appropriate titer and sex determination of bovine embryos by PCR can be feasible for confirmation.

• Korean journal of applied entomology
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• v.16 no.1 s.30
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• pp.65-67
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• 1977

Yield losses from rice dwarf virus infection are significant in Korea. Rice dwarf virus(RDV) was purified and RDV-antiserum was produced. The purified virus, mixed with an adjuvant(1:1) was injected every 10 to 14 days into rabbits. Three injections .were sufficient to produce an antiserum of 1/4,096 titer. The produced antisera will be used to facilitate the detection and identification of RDV in rice plants and in the RDV leafhopper vectors.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.1
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• pp.55-59
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• 2001

The vitellin of the red flour beetled Tribolium castaneum Herbst was purified and characterized. The vitellin of T. castaneum was purified by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In native-polyacrylamide gel electrophoresis, vitellin of T. castaneum was detected as a single band. This native vitellin has molecular weight of 440 kDa. The vitellin of T. castaneum is composed of three polypeptides, designated Vnl (178 kDa), Vn2 (168 kDa) and Vn3 (52 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph and egg extracts, but not observed in the male. These three polypeptides gradually decreased during embryogenesis. Polyclonal antiserum raised against purified vitellin reacted with the three polypeptides, Vnl, Vn2 and Vn3. Antisera raised against Vn1 and Vn2 cross-reacted with the two large subunits, Vnl and Vn2, respectively. Another subunits Vn3, however, was not cross-reacted with these two antisera. Also, antiserum raised against Vn3 did not cross-react with the Vn1 and Vn2.

• The Plant Pathology Journal
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• v.26 no.1
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• pp.25-31
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• 2010

Papaya ringspot virus (PRSV) causes the most widespread and devastating disease in papaya. Isolates of PRSV originating from different geographical regions in south India were collected and maintained on natural host papaya. The entire coat protein (CP) gene of Papaya ringspot virus-P biotype (PRSV-P) was amplified by RTPCR. The amplicon was inserted into pGEM-T vector, sequenced and sub cloned into a bacterial expression vector pRSET-A using a directional cloning strategy. The PRSV coat protein was over-expressed as a fusion protein in Escherichia coli. SDS-PAGE gel revealed that CP expressed as a ~40 kDa protein. The recombinant coat protein (rCP) fused with 6x His-tag was purified from E.coli using Ni-NTA resin. The antigenicity of the fusion protein was determined by western blot analysis using antibodies raised against purified PRSV. The purified rCP was used as an antigen to produce high titer PRSV specific polyclonal antiserum. The resulting antiserum was used to develop an immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) assay and compared its sensitivity levels with ELISA based assays for detection of PRSV isolates. IC-RT-PCR was shown to be the most sensitive test followed by dot-blot immunobinding assay (DBIA) and plate trapped ELISA.

• BMB Reports
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• v.30 no.6
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• pp.403-409
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• 1997

Studies were undertaken to develop a competitive radioimmunoassay (RIA) and indirect antigen capture enzyme-linked immunosorbent assay (ELISA) for the determination of 5'-deoxy-5'-methylthioadenosine (MTA), which is formed from decarboxylated S-adenosylmethionine by spermidine and spermine synthase. Specific antiserum against MTA was raised in rabbits by immunization with MTA-BSA which was prepared by coupling BSA to oxidized MTA with periodate. Since MTA is oxidized easily to the sulfoxide, the sulfhydryl reagent, DTT. was added to the immunogen. For RIA, immunocomplexes were separated from free MTA by using ammonium sulfate precipitation. The antiserum showed almost no cross-reactivity with a variety of other nucleotides and riboses. But, the level of cross-reactivity of 5'-isobutylthioadenosine (SIBA) was high. These results showed the importance of hydrophobicity adjacent to the 5'-OH for determining antigenicity. The lower limit of detection by this assay was 100 fmol of MTA per tube. Using this assay. MTA levels were more easily and precisely determined in biological samples when compared with HPLC analysis. The RIA procedure is less time consuming. More than 24 analyses can be carried out in 2 h and required only a very small amount of sample ($20{\mu}l$ serum). In ELISA, biotin conjugated MTA-BSA was used as the labelled MTA. The sensitivity limit of this assay was lower than 100 pmol.

• Journal of Applied Biological Chemistry
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• v.48 no.1
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• pp.16-25
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• 2005

To develop an enzyme-linked immunosorbent assay (ELISA) for the detection of the residues of the acaricide fenazaquin, five haptens were synthesized and assessed. A competitive indirect format was used with polyclonal antibodies. Under an optimized condition using the selected rabbit C antiserum, an $IC_{50}$ of $96.97\;ng{\cdot}ml^{-1}$, the detection range of $14.9{\sim}631\;ng{\cdot}ml^{-1}$, and the lowest detection limit of $8\;ng{\cdot}ml^{-1}$ were obtained. Some structurally related compounds of practical use showed low crossreactivities to the antibody. Highest cross-reactivity observed with hapten IV indicates that the antiserum C recognizes very well quinazoline ring, 4-tert-butylphenyl, and an adequate length of spacer arm. The length of spacer arm affected recognition of quinazoline ring and 4-tert-butylphenyl moieties. When applied to apple and pear, recoveries were within acceptable ranges of $93.18{\sim}104.77%$ (n = 4) and $79.40{\sim}111.95%$ (n = 4), respectively.

• Korean journal of applied entomology
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• v.16 no.2 s.31
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• pp.101-104
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• 1977

Potato virus X was purified especially for the preparation of antisera for diagnosis and identification. Potato virus X was isolated Iron infected plants by means of indicator plants and identified in electron microscopy. Isolated PVX was multiplied in tomato plants and purified by a modified procedures. The purity of PVX was 0.59mg/m1. Purified PVX was injected into rabbits once a week for 5 weeks. Antiserum was collected 10 days after the last injection. Produced antiserum was determined 1/1024 titers by means of microprecipitin tests and showed sharp reactions in agar gel-diffusion tests.

• Biomolecules & Therapeutics
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• v.17 no.1
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• pp.104-110
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• 2009

Phenylketonuria (PKU) is an inherited metabolic disorder caused by mutations in the phenylalanine catabolic enzyme, phenylalanine hydroxylase (PAH). The use of phenylalanine ammonia-lase (PAL) by oral and parenteral routes as a therapeutic drug for PKU has been severely limited due to inactivation by intestinal proteolysis and immune reactions. PEGylation was applied to PAL to reduce the degrees of antigenicity and proteolytic inactivation. Kinetic experiments with native PAL and pegylated PALs were performed, and pH stability, temperature stability, and protease susceptibility were evaluated. Enzyme linked immunosorbent assay (ELISA) was carried out to measure the immune complex between pegylated PALs and antiserum that had been extracted from a PAL-immunized mouse. Pegylated PAL, especially branched pegylated PAL (10 kDa, 1:32), was more active for phenylalanine and more stable in pancreatic proteases than native PAL. Native PAL was optimal at pH 8.5, corresponding to the average pH range of the small intestine; the same finding was noted for pegylated PALs. All linear and branched pegylated PALs had low reactivity with mouse antiserum, especially the 1:16 formulation with linear 5-kDa PEG and the 1:32 formulation with branched 10-kDa PEG. Therefore, we suggest the 1:32 formulation with branched 10-kDa PEG as the most promising formulation for enzyme replacement therapy.

• Korean Journal Plant Pathology
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• v.13 no.2
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• pp.118-124
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• 1997

The two viruses of barley yellow mosaic(BaYMV) and barley mild mosaic virus (BaMMV) were detected by ELISA from barley plants with virus-like symptoms which were collected from 16 locations in southern Korea, during 1995 and 1996. Both viruses occurred in southern Korea. Barley plants at Chongdo and Koseong were infected with BaMMV, while those infected with BaYMV were at Kurye and Taegu. After sowing 50 barley cultivars at habitually infected fields in 10 locations, the susceptibility and resistance to BaYMV and BaMMV were screened with antiserum tests. The cultivars of Albori, Alchanbori, Daejinbori, Jokangbori, Milyangbori, Boeunkwamek, Naehanssalbori, Olssalbori, Weossalbori, Dusan 29 and Deogndohyangchonkwa showed positive reaction to BaYMV antiserum, while Saeolbori, Chalbori, Jinjukwa and Baegjinkwa showed positive reaction to BaMMV. Nonsankwa 1-6 and wheat cultivars of Chongkeymil, Dahongmil, Grumil, Urimil, Jochonhomil, Sinkeyhomil showed negative reactions to both viruses. The rest cultivars were infected both with BaYMV and BaMMV. Sap inoculations to barleyplants with the two viruses of BaYMV isolated in Haenam and BaMMV isolated at National Honam Agricultural Station, expressed lower infection rate than those grown in the virus-infected fields.

• Korean journal of applied entomology
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• v.17 no.1 s.34
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• pp.29-31
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• 1978

Purely isolated cucumber mosaic virus (CMV) was multiplied in Nicotiana tabacum, Ky-57 and the virus was purified by the modified method that was developed through this study. The concentration of purified CMV was 24.25 mg/ml. The purified virus, mixed with acomplet adjuvant (1: 1) was injected into rabbits intramuscularly. Two injections at 10 day interval was enough to produce a good quality antiserum. The titer of the antiserum was 1/1280 when determined by agar gell-diffusion test. The produced antisera will be used to faciliate the detection of CMV infected vegetables and other crops.